title: Three crystal forms of the bifunctional enzyme proline utilization A (PutA) from Bradyrhizobium japonicum.
abstract: Proline utilization A proteins (PutAs) are large (1000-1300 residues) membrane-associated bifunctional flavoenzymes that catalyze the two-step oxidation of proline to glutamate by the sequential action of proline dehydrogenase and Delta(1)-pyrroline-5-carboxylate dehydrogenase domains. Here, the first successful crystallization efforts for a PutA protein are described. Three crystal forms of PutA from Bradyrhizobium japonicum are reported: apparent tetragonal, hexagonal and centered monoclinic. The apparent tetragonal and hexagonal crystals were grown in the presence of PEG 3350 and sodium formate near pH 7. The apparent tetragonal form diffracted to 2.7 A resolution and exhibited pseudo-merohedral twinning such that the true space group is P2(1)2(1)2(1) with four molecules in the asymmetric unit. The hexagonal form diffracted to 2.3 A resolution and belonged to space group P6(2)22 with one molecule in the asymmetric unit. Centered monoclinic crystals were grown in ammonium sulfate, diffracted to 2.3 A resolution and had two molecules in the asymmetric unit. Removing the histidine tag was important in order to obtain the C2 crystal form.
authors.affiliation: Department of Chemistry, University of Missouri-Columbia, Columbia, MO 65211, USA.
authors.forename: Jonathan P
authors.initials: JP
authors.lastname: Schuermann
authors.forename: Tommi A
authors.initials: TA
authors.lastname: White
authors.forename: Dhiraj
authors.initials: D
authors.lastname: Srivastava
authors.forename: Dale B
authors.initials: DB
authors.lastname: Karr
authors.forename: John J
authors.initials: JJ
authors.lastname: Tanner
mesh_terms: D019747:Aldose-Ketose Isomerases / Q000737:chemistry* / Q000378:metabolism; D019843:Archaeal Proteins / Q000737:chemistry* / Q000378:metabolism; D020369:Bradyrhizobium / Q000201:enzymology*; D018360:Crystallography, X-Ray; D011392:Proline / Q000378:metabolism*; D011487:Protein Conformation
chemical_list: D019843:Archaeal Proteins; D011392:Proline; D019747:Aldose-Ketose Isomerases; C019714:D-arabinose isomerase